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@ARTICLE{Polverini:51048,
author = {Polverini, E. and Cugini, G. and Annoni, F. and
Abbruzzetti, S. and Viappiani, C. and Gensch, T.},
title = {{M}olten globule formation in apomyoglobin monitored by the
fluorescent probe nile red},
journal = {Biochemistry},
volume = {45},
issn = {0006-2960},
address = {Columbus, Ohio},
publisher = {American Chemical Society},
reportid = {PreJuSER-51048},
pages = {5111 - 5121},
year = {2006},
note = {Record converted from VDB: 12.11.2012},
abstract = {The interaction of nile red (NR) with apomyoglobin (ApoMb)
in the native (pH 7) and molten globule (pH 4) states was
investigated using experimental and computational methods.
NR binds to hydrophobic locations in ApoMb with higher
affinity (K(d) = 25 +/- 5 microM) in the native state than
in the molten globule state (K(d) = 52 +/- 5 microM). In the
molten globule state, NR is located in a more hydrophobic
environment. The dye does not bind to the holoprotein,
suggesting that the binding site is located at the heme
pocket. In addition to monitoring steady-state properties,
the fluorescence emission of NR is capable of tracking
submillisecond, time-resolved structural rearrangements of
the protein, induced by a nanosecond pH jump. Molecular
dynamics simulations were run on ApoMb at neutral pH and at
pH 4. The structure obtained for the molten globule state is
consistent with the experimentally available structural
data. The docking of NR with the crystal structure shows
that the ligand binds into the binding pocket of the heme
group, with an orientation bringing the planar ring system
of NR to overlap with the position of two of the heme
porphyrin rings in Mb. The docking of NR with the ApoMb
structure at pH 4 shows that the dye binds to the heme
pocket with a slightly less favorable binding energy, in
keeping with the experimental K(d) value. Under these
conditions, NR is positioned in a different orientation,
reaching a more hydrophobic environment in agreement with
the spectroscopic data.},
keywords = {Animals / Apoproteins: chemistry / Apoproteins: metabolism
/ Binding Sites / Computer Simulation / Fluorescent Dyes:
analysis / Horses / Hydrogen-Ion Concentration / Models,
Molecular / Myoglobin: chemistry / Myoglobin: metabolism /
Oxazines: analysis / Oxazines: chemistry / Protein Binding /
Protein Folding / Protein Structure, Tertiary / Apoproteins
(NLM Chemicals) / Fluorescent Dyes (NLM Chemicals) /
Myoglobin (NLM Chemicals) / Oxazines (NLM Chemicals) /
apomyoglobin (NLM Chemicals) / nile red (NLM Chemicals) / J
(WoSType)},
cin = {IBI-1},
ddc = {570},
cid = {I:(DE-Juel1)VDB57},
pnm = {Funktion und Dysfunktion des Nervensystems},
pid = {G:(DE-Juel1)FUEK409},
shelfmark = {Biochemistry $\&$ Molecular Biology},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:16618100},
UT = {WOS:000237049400004},
doi = {10.1021/bi051905y},
url = {https://juser.fz-juelich.de/record/51048},
}